HIV/AIDS is a priority public health condition. There are about 1.1 million HIV infected people in the USA and 56,300 new HIV infections annually. The critical barrier to slowing the spread of HIV and treating AIDS is the lack of a rapid simple test for guiding anti-retroviral treatment (ART) in point-of-care (POC) settings. Low- resource POC community clinics are the largest provider of HIV healthcare in the USA. The existing tests for the therapeutic monitoring of AIDS are the CD4 T cell count (CD4) and the HIV-RNA viral load (VL). These tests are invasive, expensive, take several days and require blood work equipment and costly laboratory infrastructure. These resources are not available in POC. The ultimate goal of this project is to develop a rapid oral test for therapeutic monitoring of HIV/AIDS patients in POC. The test will measure saliva-based Stress Response Profiling (SRP) biomarkers using a low-cost, simple device, the lateral-flow immunoassay (LFIA). Preliminary Studies (n=17) showed that 40 SRP biomarkers were >20-fold increased in ART-unresponsive AIDS patients and returned to the normal baseline in ART-responsive AIDS patients. The effect was clinically specific because this panel of SRP biomarkers was not increased by non-HIV infections or diseases. The objective of the proposed research it the proof-of-concept for the proposed product. Aims 1-4 will validate a SRP biomarker panel with diagnostic accuracy and clinical specificity, and translate selected biomarkers into a prototype LFIA device. The clinical collaborator on the project is the AIDS Healthcare Foundation (AHF). AHF is the largest nonprofit provider of HIV healthcare in the USA, through a network of POC community clinics. In Aim 1, saliva samples will be collected from 3 groups of AHF patients (n=72): ART-responsive and ART-unresponsive AIDS patients, and HIV-negative subjects. In Aim 2, 40 SRP biomarkers selected in Preliminary Studies will be measured in the clinical saliva samples using immunohistochemistry and automated image analysis, a highly sensitive, specific and reliable assay. Candidate biomarkers with e 2-fold higher measurements in ART-unresponsive AIDS patients will be confirmed using Western blots. The best 10 candidate markers will be measured using ELISA assays in Aim 3. The ELISA measurements will be analyzed using ROC plots to determine diagnostic accuracy and clinical specificity of the biomarkers.In Aim 4, 3 SRP biomarkers with the best diagnostic accuracy and specificity will be optimized into a prototype LFIA device. ELISA assays validated in Aim 3 will serve as reference assays during the LFIA development. The proposed diagnostic test is noninvasive, rapid and cost-effective. It will radically improve clinical HIV/AIDS care through significantly improved deliverability and acceptability of therapeutic monitoring in POC. The new test is highly innovative and has strong commercial potential because there is no existing POC test for therapeutic monitoring of AIDS. In addition to the direct impact on HIV healthcare, the proposed introduction of salivary biomarkers for therapeutic monitoring of HIV/AIDS will also greatly expand the scope of salivary diagnostics. PUBLIC HEALTH RELEVANCE: HIV/AIDS is a priority public health condition in the USA. The major challenge in treating HIV-infected people early and effectively is the lack of a rapid simple test that could be used to guide the treatment in low-resource community clinics. The goal of this project is to develop a noninvasive hand-held test for guiding AIDS treatment. The proposed product has potential to bring significant benefits for public health by providing a pain- free, low-cost and easy-to-use alternative to existing tests.